New Kit-based Solutions Provide Highly Flexible Microrna Library Creation

Integrated DNA Technologies (IDT) has announced the availability of its new proprietary miRCat? small RNA cloning system. The unique kit-based resources make the creation of small RNA (including miRNAs, piRNAs and endogenous siRNAs) libraries from any primary RNA source, highly time and cost efficient with peerless flexibility and consistency. The new miRCat system permits cloning of very rare small RNAs and takes into account the natural variability in structure and sequence between species. Conveniently, it is compatible with most existing standard laboratory protocols for processes such as RNA extraction, purification and cloning. These features make the miRCat small RNA cloning system the ideal solution for all small RNA research.

Based upon a pre-activated adenylated oligonucleotide linkering method, the miRCat cloning system has been carefully designed to make small RNA library creation easy for all researchers. It consists of three sequential protocols: RNA isolation and enrichment, followed by cloning linker attachment, and ending with an amplification and cloning phase. The system can be used for concatamer cloning or direct ?shotgun' cloning; it also can provide PCR amplicons suitable for use with TOPO TA Cloning® or pGEM® T-Easy cloning vectors.

For cloning small RNAs lacking the 5'-phosphorylated end present in miRNAs, a modified kit - miRCat-33? - has been developed, which uses 5' ligation-independent cloning. This makes it an excellent cloning system for the recently discovered C. elegans 5' tri-phosphorylated microRNAs, which would otherwise be omitted. Furthermore, optional kit components ensure additional versatility with a choice of three different 3' cloning linkers. These provide various restriction site combinations to fit with any preferred cloning processes. Specific primers are also available to convert the resultant small RNA libraries using Roche 454 Life Sciences sequencing systems.